We next addressed whether cleaved amyloid β peptides localize similarly to APP using a peptide-specific antibody.
To this end, we carried out a peptide spot analysis using a polypeptide library presenting 1,849 distinct C termini prepared by an inverted peptide array (Boisguerin et al., 2007).
Intensities were calculated using a grid with 192 circular features of 2 mm diameter, each centered around a peptide spot to avoid scoring SPOTs with halo or rings.
Peptides were synthesized on cellulose membranes using a MultiPep SPOT peptide arrayer (Intavis).
Membranes Synthesis Peptides for the first two arrays were synthesized directly on cellulose membranes (Intavis) using a MultiPep SPOT peptide arrayer (Intavis) and commercially available standard (Intavis) or modified (Bachem) L-amino acid precursors as previously described (Nady et al., 2008 ).
Using peptide SPOT arrays, we identified the minimal binding epitope to be EKLMKAFESLKSFQ, comprising the N-terminal residues 5 18 of Htt and including the first residue of the poly-Gln stretch.
To characterize the myosin-binding canyon, we carried out systematic peptide binding studies using ITC and peptide spot analyses.
The urinary C-peptide creatinine ratio (UC-peptide creatininedescratio method of assessing C-peptide prodUCPCRn that isvolves using a spot urecentlyt.
Briefly, each protein spot on SYPRO Ruby stained gels was picked using a spot picker (Amersham Biosciences).
Characterization of prorenin activation using a synthetic peptide substrate.
